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WO2010118034A2 - Analogues peptidiques cycliques utilisés en imagerie non invasive des cellules bêta du pancréas - Google Patents

Analogues peptidiques cycliques utilisés en imagerie non invasive des cellules bêta du pancréas Download PDF

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WO2010118034A2
WO2010118034A2 PCT/US2010/030103 US2010030103W WO2010118034A2 WO 2010118034 A2 WO2010118034 A2 WO 2010118034A2 US 2010030103 W US2010030103 W US 2010030103W WO 2010118034 A2 WO2010118034 A2 WO 2010118034A2
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lys
glu
glp
seq
imaging
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WO2010118034A9 (fr
WO2010118034A3 (fr
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Jung-Mo Ahn
Xiankai Sun
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University of Texas System
University of Texas at Austin
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University of Texas System
University of Texas at Austin
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics

Definitions

  • the present invention relates in general to the field of imaging, and more particularly, to novel cyclic glucagon-like peptide and Exendin analogues for non-invasive imaging of pancreatic beta-cells to diagnose, e.g., diabetes mellitus.
  • Diabetes mellitus is a chronic disease characterized by multiple metabolic abnormalities resulting in impaired management of glucose. According to the recent statistics, diabetes is the fifth leading cause of death in the United States. Diabetic patients are also at significantly higher risk to develop complications which severely influence life quality of the patients.
  • Type 1 diabetes A hallmark of Diabetes is high level of blood glucose caused by the lack of insulin production, insulin resistance in peripheral tissues, or both, and generally classified into two types, insulin- dependent (type 1) and non-insulin-dependent (type 2).
  • Type 1 diabetes is found to be connected with the loss of pancreatic beta-cells which secretes insulin upon feeding.
  • pancreatic beta-cells which secretes insulin upon feeding.
  • the present invention includes novel cyclic glucagon- like peptide (GLP-I) or Exendin analogues used to assess pancreatic beta-cells using non-invasive imaging techniques.
  • GLP-I sequence includes H 7 AEGT FTSDV SSYLE GQAAK EFIAW LVKGR 36 SEQ ID No: 1 which is a 30 amino acid-containing peptide that is produced by intestinal L-cells.
  • the Exendin-4 sequence includes His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu- Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro- Ser-NH2 (SEQ ID No: 2).
  • the cyclic analog comprises a portion of a peptide or protein that binds specifically to the GLP-I receptor (GLP-IR) and the cyclic analog has one or more conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, salts and derivatives thereof wherein the cyclic analog is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live.
  • GLP-IR GLP-I receptor
  • the present invention includes novel cyclic GLP-I analogues used to assess pancreatic beta- cells using non-invasive imaging techniques.
  • GLP-I is an endogenous hormone that is known to interact with a receptor on the pancreatic beta-cells. However, rapid enzymatic degradation of this peptide in vivo prevents its effective use.
  • the novel cyclic GLP-I analogues are extremely stable against enzymes that are known to participate in the GLP-I degradation. In addition, these cyclic GLP-I analogues were found to have higher potency when compared to the native GLP-I.
  • PET positron emission tomography
  • the present invention includes composition, methods and agents comprising at least portions of GLP-I with one or more conformational restrictions, including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, between the positions 7 and 36 of GLP-I, salts and derivatives thereof wherein the agent is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half-live.
  • conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, between the positions 7 and 36 of GLP-I, salts and derivatives thereof wherein the agent is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half-live.
  • the agent is selected from at least one of: c[Glu 16 ,Lys 20 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, c[Glu 16 ,Lys 2O ]-c[Glu 3 °,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1; c[Glu 18 ,Lys 22 ]- c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, [D-Ala 8 ]-c[Glu 16 ,Lys 20 ]GLP-l(7-36)- NH 2 SEQ ID No: 1, [D-Ala8]-c[Glu30,Lys34]GLP- 1(7-36)- NH 2 SEQ ID No: 1, [D-AIa 8 ]-
  • the agent binds specifically to the GLP-I receptor.
  • the agent is multivalent.
  • the agent further comprises at least one of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of 18 F,
  • the agent further comprises at least one of a therapeutic or cytotoxic agent.
  • the agent further comprises at least one of an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • the imaging agent binds specifically to pancreatic tissue.
  • the imaging agent further comprises a pharmaceutically acceptable excipient.
  • the imaging agent is formulated for use in a diagnostic method practiced on the human or animal body.
  • the imaging agent has an increased resistance to proteolytic cleavage by dipeptidyl peptidase-IV (DPP-IV), neutral endopeptidase (NEP), or both.
  • DPP-IV dipeptidyl peptidase-IV
  • NEP neutral endopeptidase
  • the imaging agent is an organ specific imaging agent comprises one or more labels that made the agent detectable by positron emission tomography (PET), single photon emission computed tomography (SPECT), radioscintigraphy, or magnetic resonance imaging (MRI).
  • PET positron emission tomography
  • SPECT single photon emission computed tomography
  • MRI magnetic resonance imaging
  • the imaging agent is adapted for imaging pancreatic beta cells.
  • the present invention includes composition, methods and agents for imaging a pancreas comprising: injecting into a patient in need of pancreatic imaging an effective amount of a contract agent comprising at least a portion of GLP-I with one or more conformational restrictions, including but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, between the positions 7 and 36 of GLP-I, salts and derivatives thereof, wherein the agent is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live.
  • the agent is selected from at least one of: c[Glu 16 ,Lys 20 ]GLP-l(7-36)-NH 2
  • the agent binds specifically to the GLP-I receptor.
  • the agent is multivalent.
  • the agent further comprises at least one of a radio label, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of 18 F, 68 Ga, 60/61/62/64 c u , 89 Zr, 86 Y, 124 I, 99m Tc, 94m Tc, 111 In, 67 Ga, 125 I, 123 I, 177 Lu, 75/76/77 Br, 166 Ho, and 153 Sm.
  • the agent further comprises at least one of a therapeutic or cytotoxic agent.
  • the agent further comprises at least one of an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • the imaging agent binds specifically to pancreatic tissue.
  • the imaging agent further comprises a pharmaceutically acceptable excipient.
  • the imaging agent is formulated for use in a diagnostic method practiced on the human or animal body.
  • the imaging agent has an increased resistance to proteolytic cleavage by dipeptidyl peptidase-IV (DPP-IV), neutral endopeptidase (NEP), or both.
  • the imaging agent is an organ specific imaging agent comprises one or more labels that made the agent detectable by positron emission tomography (PET), single photon emission computed tomography (SPECT), radioscintigraphy, or magnetic resonance imaging (MRI).
  • the present invention includes an imaging agent comprising at least one of: c[Glu 16 ,Lys 20 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, c[Glu 16 ,Lys 2O ]-c[Glu 3 °,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1; c[Glu 18 ,Lys 22 ]- c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, [D-Ala 8 ]-c[Glu 16 ,Lys 20 ]GLP-l(7-36)- NH 2 SEQ ID No: 1, [D-Ala8]-c[Glu30,Lys34]GLP- 1(7-36)- NH 2 SEQ ID No: 1, [D-AIa 8
  • the agent binds specifically to the GLP-I receptor.
  • the agent is multivalent.
  • the agent further comprises at least one of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of 18 F, 68 Ga, 60/61/62/64 c u , 89 Zr, 86 Y, 124 I, 99m Tc, 94m Tc, 111 In, 67 Ga, 125 I, 123 I, 177 Lu, 75/76/77 Br, 166 Ho, and 153 Sm.
  • the agent further comprises at least one of a therapeutic or cytotoxic agent, e.g., an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an antimitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • a therapeutic or cytotoxic agent e.g., an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an antimitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • the imaging agent binds specifically to pancreatic tissue.
  • the imaging agent further comprises a pharmaceutically acceptable excipient.
  • the imaging agent is formulated for use in a diagnostic method practiced on the human or animal body.
  • the imaging agent has an increased resistance to proteolytic cleavage by dipeptidyl peptidase-IV (DPP-IV), neutral endopeptidase (NEP), or both.
  • the imaging agent is an organ specific imaging agent comprises one or more labels that made the agent detectable by positron emission tomography (PET), single photon emission computed tomography (SPECT), radioscintigraphy, or magnetic resonance imaging (MRI).
  • One embodiment of the present invention includes a multivalent GLP-I having an optionally substituted multivalent composition conjugated to two or more GLP-I molecules to form the multivalent GLP-I, wherein the multivalent GLP-I is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live.
  • the present invention includes a diagnostic or imaging agent comprising [D-Ala 8 ]-c[Glu 18 ,Lys 22 ]-c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1, salts or derivatives thereof.
  • the agent is multivalent.
  • the present invention includes a composition having an agent comprising at least portions of an Exendin-4 protein having one or more conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, salts and derivatives thereof wherein the agent is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live.
  • an agent comprising at least portions of an Exendin-4 protein having one or more conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, salts and derivatives thereof wherein the agent is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live.
  • the present invention includes a diagnostic or imaging agent having at least one of: a cyclic analog imaging agent comprising a portion of a peptide or protein that binds specifically to the GLP-I receptor (GLP-IR) and the cyclic analog has one or more conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, salts and derivatives thereof wherein the cyclic analog is more stable than a non- cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live, wherein the cyclic analog comprises at least a portion of a GLP-I peptide or at least a portion of an Exendin-4 peptide salts, derivatives or combinations thereof.
  • GLP-IR GLP-I receptor
  • the agent is selected from at least one of: c[Glu lo ,Lys 14 ]Exendin-4(l-39)-NH2 SEQ ID No: 2, c[Glu 24 ,Lys 28 ]Exendin-4(l-39)-NH 2 SEQ ID No: 2, c[Glu 12 ,Lys 16 ]Exendin-4(l- 39)- NH 2 SEQ ID No: 2, c[Glu lo ,Lys 14 ]-c[Glu 24 ,Lys 28 ]Exendin-4(l-39)- NH 2 SEQ ID No: 2; c[Glu 12 ,Lys 16 ]-c[Glu 24 ,Lys 28 ]Exendin-4(l-39)-NH 2 SEQ ID No: 2, [Aib 8 ]- c[Glu lo ,Lys 14 ]Exendin-4(l-39)-NH 2 SEQ ID No: 2, [Aib 8 ]-c[Glu lo ,Lys 14 ]Ex
  • the agent binds specifically to the GLP-I receptor.
  • the agent is multivalent.
  • the agent further comprises at least one of a radio label, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of 18 F, 68 Ga, 60/61/62/6 4Cu, 89 Zr, 86 Y, 124 I, "mTc, 111 In, 67 Ga, 125 I, 123 I, 177 Lu, 166 Ho, and 153 Sm.
  • the agent further comprises at least one of a therapeutic or cytotoxic agent.
  • Figure 1 is a diagram that shows the introduction of a lactam bridge to GLP-I, and the enhanced binding obtained with the bicyclic GLP-I analogues of the present invention.
  • Figure 2 shows the in vitro specific binding of GLP- 1(7-36)- NH 2 (left) SEQ ID No: 1 and [D- Ala8]-GLP- 1(7-36)- NH 2 (right) SEQ ID No: 1 to INS-I embedded collagen beads.
  • Upper panel autoradiography images; lower panel: semi-quantitation of the autoradiography images.
  • Figure 3 is a chart that shows receptor activation by cyclic GLP-I analogues
  • Figure 4 are HPLC chromatograms of a bicyclic GLP-I analogue after incubation with DPP-IV and NEP, Green, GLP-I (7-36)-NH 2 (3 h) SEQ ID No: 1; Blue, GLP-I (7-36)-NH 2 (24 h) SEQ ID No: 1, Red, [D-Ala 8 ]-c[Glu 18 ,Lys 22 ]-c[Glu 30 ,Lys 34 ]GLP-l(7-36)-NH 2 (24 h) (EM2198) SEQ ID No: 1.
  • Figure 5 is the Synthesis of a PET imaging agent by using a bicyclic GLP-I analogue.
  • Figure 6 is a graph that shows the binding of L-GLP-I (GLP-I (7-36)-NH2) SEQ ID No: 1, D- GLP-I ([D-Ala8]GLP- 1(7-36)- NH 2 ) SEQ ID No: 1, EM2196 ([D-Ala8]-c[Glul6,Lys20]- c[Glu30,Lys34]GLP- 1(7-36)- NH 2 SEQ ID No: 1) and EM2198 ([D-Ala8]-c[Glul8,Lys22]- c[Glu30,Lys34]GLP- 1(7-36)- NH 2 SEQ ID No: 1) to various organs.
  • Figure 7 is a PET image of the binding of EM2198 in a mouse.
  • Figure 8 is a PET image of the binding of EM2198 in a mouse with blocking with Exendin-4.
  • Figure 9 are graphs that show a comparison of contrast in pancreatic, liver and kidney microPET scans.
  • Figure 10 are transaxial PET/CT images at 30 min p.i. of EM2198 with or without exendin-4 blocking.
  • Figure 11 is a graph that shows that EM2198 specifically target the GLP-IR in pancreas.
  • Figure 12 is a graph that illustrates binding of the Exendin-4 to target GLP-I receptors. Description of the Invention
  • novel cyclic GLP-I analogues can be labeled with proteins, radionuclides, fluorescent labels, metals, chromogenic agents, enzymes and other agents that enhace its use as an imaging agent ,s.
  • the imaging agent further comprises at least one of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti- angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • the imaging agent may include or more labels that make the agent detectable by positron emission tomography (PET), single photon emission computed tomography (SPECT), radioscintigraphy, magnetic resonance imaging (MRI) and computed tomography (CT scan).
  • the agents disclosed herein have been found to have increased resistance to proteolytic cleavage by dipeptidyl peptidase-IV (DPP-IV), neutral endopeptidase (NEP), or both.
  • DPP-IV dipeptidyl peptidase-IV
  • NEP neutral endopeptidase
  • GLP-I glucagon- like peptide- 1
  • it may be an ideal candidate to be employed for non-invasive imaging of the beta-cells.
  • its rapid metabolism hampers the use of GLP-I as in vivo imaging agents.
  • the limited number of GLP-I receptors on the beta-cells requires high specificity and sensitivity for imaging studies.
  • MRI magnetic resonance imaging
  • CT computed tomography
  • pancreas is a highly vascularized soft organ and the islets only represent 2-3 % of the pancreatic tissues.
  • imaging or contrast agents that recognize the scarcely dispensed beta-cells within pancreas and are responsive to their biological functions, must be developed.
  • tomographic nuclear imaging approaches especially positron emission tomography (PET) have demonstrated their significant importance and promising potential in applications of molecular imaging probes due to the superior sensitivity and specificity in diverse subjects, and the ability to quantitatively analyze the regions of interest (2-5).
  • PET positron emission tomography
  • PET imaging methods have gained a considerable momentum to move forward to the molecular imaging of the pancreatic beta-cells.
  • PET imaging techniques can be mainly defined by the successful development of radiotracers that specifically target the pancreatic beta-cells.
  • Monoclonal antibodies and peptides which are specific to cell membrane receptors have been used as targeting molecules for cancer diagnosis and therapy (6, 7).
  • peptides have shown more efficient tissue penetration and rapid clearance from non- target organs, and normally are not immunogenic upon repetitive administration.
  • glucagon-like peptide- 1 plays a critical role in the function of beta-cells.
  • GLP-I (sequence: H 7 AEGT FTSDV SSYLE GQAAK EFIAW LVKGR 36 )
  • SEQ ID No: 1 is a 30 amino acid-containing peptide that is produced by intestinal L-cells. Its predominant bioactive form is GLP- 1(7-36) amide, which is considered as the endogenous ligand to the GLP-I receptor (S).
  • GLP- 1 is secreted from intestinal L-cells into the blood stream (8-10).
  • the circulating hormone acts on pancreatic beta-cell GLP-I receptors, and enhances glucose-stimulated insulin release In addition to its insulinotropic effects, GLP-I also limits postprandial glucose elevation through several other mechanisms, including (1) stimulation of beta-cell growth and survival (13,14); (2) inhibition of glucagon release from pancreatic alpha-cells (15); (3) delay of gastric emptying via vagal mechanisms (16,17); and (4) inhibition of short-term food intake by modulation of neuronal activity in the brain (18,19).
  • GLP-I receptor is a seven transmembrane-spanning G-protein coupled receptor (GPCR), and upon ligand-binding GLP-IR undergo conformational change which leads to the production of secondary messengers including cAMP and Ca 2+ for its physiological functions.
  • GPCR G-protein coupled receptor
  • the size of GLP-IR is large (463 residues) and it employs a long N-terminal chain (120 residues) and large extracellular loops to accommodate its large peptide ligand (20).
  • GLP-I interacts with its receptor with high binding affinity (K d ⁇ 1 nM) and directly involves in the function of the beta-cells, it appears to be a suitable candidate for beta-cell imaging.
  • GLP-I is highly susceptible to proteases including ubiquitous dipeptidyl-peptidase IV (DPP-IV). This enzyme cleaves two residues (His 7 -Ala 8 ) from the N-terminus of GLP-I, which are highly important for both of receptor binding and activation. The degradation by DPP-IV results in a fragment, GLP- 1(9-36) amide, which lost receptor affinity and biological activity nearly completely.
  • DPP-IV ubiquitous dipeptidyl-peptidase IV
  • GLP-I, D-GLP-I (D-alanine at the 8 th position), and two bicyclic GLP-I peptide analogues were synthesized by Fmoc solid-phase chemistry, followed by the coupling of l,4,7,10-Tetraazacyclododecane-l,4,7-tris-acetic acid-10-maleimidoethyl- acetamide (maleimido-mono-amide-DOTA? to the Cys at the C-terminus of the peptides.
  • the peptide conjugates were labeled with 64 Cu under a mild condition.
  • the in vitro stability of the 64 Cu-labeled peptides was evaluated in rat serum at 37 0 C. After protein precipitation with ethanol, the serum mixture was centrifuged and the supernatant was analyzed by radio-HPLC. The half maximal effective concentrations (EC50) of the peptides were determined by the dose- response of the peptide triggered cyclic AMP (cAMP) accumulation using HEK293 cells, which stably express GLP-IR-GFP in the presence of the phosphodiesterase inhibitor 3-isobutyl-l- methylxanthine (IBMX) at 37 0 C.
  • cAMP cyclic AMP
  • the peptide conjugates were synthesized and characterized by HPLC and MALDI-Mass spectroscopy. The specific activity of the labeled peptides was up to 6.0 x 10 5 Ci/mol with radiochemical purity > 99%. After 1 h incubation in rat serum, all peptides showed similar stability ( ⁇ 50 %) as determined by radio-HPLC.
  • both bicyclic peptides showed markedly higher agonistic potency in triggering cAMP accumulation (EC 50 of EM2196 and EM2198: ⁇ 1 nM; EC 50 of GLP- 1 : 5 nM).
  • Dynamic PET-CT imaging revealed rapid pancreas uptake ( ⁇ 5 min) and high renal accumulation Of 64 Cu labeled peptides in normal mice. While GLP-I, D-GLP-I, and EM2196 showed fast clearance ( ⁇ 10 min) from the pancreas, EM2198 demonstrated significantly longer pancreatic retention (> 30 min). Post-PET biodistribution data was in agreement with the imaging findings.
  • FIG. 1 is a diagram that shows the introduction of a lactam bridge to GLP-I, and the enhanced binding obtained with the bicyclic GLP-I analogues of the present invention. Twelve cyclic GLP-I analogues containing lactam bridges in various regions (from the N- to C-terminal regions) were designed and synthesized.
  • the cyclic peptides were prepared by following standard solid-phase peptide synthesis protocol with N-Fmoc/ l Bu protecting group strategy.
  • the side chains of a lysine and a glutamic acid that would form a lactam bridge were protected with allyl groups which were selectively removed by using Pd 0 catalyst and an allyl scavenger like N,N-dimethylbarbituric acid (29).
  • the released free amine and carboxylic acid were coupled on resin using HBTU or BOP to form a cyclic peptide.
  • the prepared cyclic GLP-I analogues were assessed for their receptor-binding and activation using HEK293 cells stably expressing human GLP-I receptors, and competitive receptor binding assay of the peptides was carried out using 125 I-exendin(9-39) as a radioligand in the presence of a DPP-IV inhibitor (30).
  • cAMP formation by the peptides was determined by radioimmunoassay using the transfected HEK293 cells to examine agonistic activity.
  • cyclic GLP-I analogues Among the twelve cyclic GLP-I analogues, six peptides showed comparable or improved receptor-binding and activation, such as c[Lys 16 ,Glu 20 ]GLP-l(7-36)-NH 2 (SEQ ID No: 1) and c[Lys 30 ,Glu 34 ]GLP-l(7-36)-NH 2 SEQ ID No: 1. These cyclic GLP-I analogues also showed enhanced stability against enzymatic degradation when assessed with isolated enzymes (DPP-IV and NEP 24.11) and kidney cells over 24 h.
  • DPP-IV and NEP 24.11 isolated enzymes
  • GLP-I receptor (GLP-IR) is mainly expressed on pancreatic beta-cells but to a less extent by lung, heart, kidneys, gastrointestinal tract, or brain. While GLP-I analogues have been extensively studied for the treatment of type
  • GLP-I analogues were evaluated by in vitro binding assay and in vivo tissue distribution.
  • a linear GLP-I analogue, [Lys(Ac) 16 ,Gln 20 ]GLP-l(7-36)-NH 2 , SEQ ID No: 1 was labeled with 125 I (at Tyr 19 ).
  • This peptide showed an appreciable specific binding to freshly isolated rat islets as determined by a displacement binding assay, the tissue distribution of the peptide in normal Sprague-Dawley rats demonstrated no significant pancreas uptake and instead high accumulation in kidneys and stomach. This presumably resulted from the degradation of the peptide by DPP-IV since it has Ala at position 8.
  • DOTA 1,4,7,10- tetraazacyclododecane-l,4,7,10-tetraacetic acid
  • Standard thiol-maleimide conjugation provided coupling of DOTA to both L-GLP-I ([Ala 8 ]GLP-l-Ahx-Cys) SEQ ID No: 1 and D-GLP-I ([D-Ala 8 ]GLP-l- Ahx-Cys) SEQ ID No:l, and the DOTA-GLPl conjugates were characterized and purified using HPLC and MALDI-MS. The purified conjugates were labeled with either 64 Cu or 111 In in high radiochemical yields.
  • Scheme 1 Conjugation of DOTA-maleimide with GLP-I peptides
  • Figure 2 shows the in vitro specific binding of L-GLP-I (left) and D-GLP-I (right) to INS-I embedded collagen beads.
  • Upper panel autoradiography images; lower panel: semi-quantitation of the autoradiography images.
  • INS-I an insulinoma cell line
  • embedded collagen beads -1,000 cells/bead
  • biodistribution results revealed the peptide accumulation only in the pancreas.
  • Fig. 2 it is reasonable to assume that the elevated pancreas uptake of the D-GLP-I conjugate resulted from the specific beta-cell targeting.
  • islets of Langerhans were isolated post-biodistribution or PET imaging.
  • Enzymatically stable GLP-I analogues The stability of GLP-I analogues affects in vivo imaging.
  • a series of cyclic GLP-I analogues were designed and synthesized that include lactam bridges between Lys 1 and GIu 1+4 and which stabilize the ⁇ - helical structure in GLP-I (28).
  • lactam bridges between Lys 1 and GIu 1+4 and which stabilize the ⁇ - helical structure in GLP-I (28).
  • a number of cyclic peptides were found to have comparable or improved receptor-binding and activation, and showed moderate enzymatic stability when incubated with kidney cells.
  • cyclic peptides are: c[Lys 16 ,Glu 20 ]GLP-l(7- 36)-NH 2 (1C) SEQ ID No: 1, c[Lys 18 ,Glu 22 ]GLP-l(7-36)-NH 2 (2C) SEQ ID No: 1, c[Lys 22 ,Glu 26 ]GLP-l(7-36)-NH 2 (3C) SEQ ID No: 1, c[Lys 23 ,Glu 27 ]GLP-l(7-36)-NH 2 (4C) SEQ ID No: 1, c[Lys 26 ,Glu 30 ]GLP-l(7-36)-NH 2 (5C) SEQ ID No: 1, and c[Lys 30 ,Glu 34 ]GLP- l(7-36)-NH 2 (6C) SEQ ID No: 1.
  • DPP-IV is not the only enzyme responsible for the metabolism of GLP-I.
  • neutral endopeptidase cleaves multiple sites in GLP-I and the cleavage sites are Asp 15 -Val 16 , Ser 18 -Tyr 19 , Tyr 19 -Leu 20 , Glu 27 -Phe 28 , Phe 28 -Ile 29 , and Trp 31 -Leu 32 (32). Since all of these sites are within the sequences stabilized by the lactam bridges in 1C-6C, the cyclization via the lactam bridge provides sufficient stabilization over NEP 24.11.
  • bicyclic GLP-I analogues were synthesizes that protect against NEP 24.11.
  • bicyclic GLP- 1 analogues BC1-BC9 were synthesized using the standard solid-phase peptide synthesis technique as successfully used in the preparation of the cyclic GLP-I analogues.
  • Each of the bicyclic peptides has D-AIa 8 for DPP-IV stability and a cysteine at the C-terminus for conjugating to a multivalent metal chelator via an aminohexanoic acid as a spacer.
  • the bicyclic structure and D-AIa 8 substitution provided substantial enzymatic stability to facilitate beta-cell imaging.
  • a linear peptide of which sequence contains only the first lactam bridge was grown on solid support using Fmoc-protected amino acids.
  • allyl protecting groups on a Lys and a GIu that are to form the first lactam bridge were removed selectively and the released free amine and carboxylic acid were coupled to form a monocyclic peptide.
  • the percent injected dose per gram (%ID/g) and percent injected dose per organ (%ID/organ) was calculated by comparison to a weighed, counted standard. Pharmacokinetic parameters were estimated by using either one- or two- compartment model. Specifically, blood (10-20 ⁇ L) was drawn from the animals at 5, 10, 20, and 40 min, and counted on a gamma counter.
  • Urine excretion experiments were carried out with the last time point animal groups. Rat urine was collected from metabolic cages at selected time points post-injection. In addition, the urine was concentrated under nitrogen so that radio-TLC or radio-HPLC analysis can be performed to determine the percentage of intact peptide conjugates.
  • Type 1 diabetic rats were established by administering streptozotocin (STZ; 65 mg/kg in 20 mM citrate saline buffer) intraperitoneally (i.p.) to mature Sprague-Dawley rats fasted for two days (40-42).
  • STZ streptozotocin
  • the biodistribution procedures were the same as above, but only one time point that showed the highest pancreas uptake was selected. Histomorphometry of pancreas and islets were performed after the dissection to determine the percentage of beta-cell fraction in the rat's pancreas with established methods (43-46).
  • PET imaging evaluation of selected 64 Cu-labeled GLP-I constructs in diabetic rats Many GLP- 1 constructs with a positive pancreas uptake and beta-cell density correlation were evaluated using PET imaging.
  • Type 1 diabetes rat model were used: The biobreeding diabetes-prone (BB- DP) rats available from Biomedical Research Models Inc. (Worcester, MA) spontaneously develop type 1 diabetes with an average onset of hyperglycemia at 10 weeks of age. Blood glucose levels and the area under the curves of the intraperitoneal glucose tolerance test (AUC IPGTT) were monitored daily and twice a week from the rat's age of 50 days, respectively, by established methods (46-48).
  • BB- DP biobreeding diabetes-prone rats available from Biomedical Research Models Inc.
  • AUC IPGTT intraperitoneal glucose tolerance test
  • a series of cyclic GLP-I analogues to determine the receptor-bound conformation of GLP-I was employed to stabilize an ⁇ -helical structure in various regions of GLP-I and was demonstrated that a strategically positioned conformational restriction can enhance interaction with the receptor.
  • Bicyclic GLP-I analogues were synthesized by using standard Fmoc/t-butyl solid-phase peptide chemistry.
  • GIu(OAl) and Lys(Aloc) were introduced in a linear peptide and the allyl protecting groups were selectively removed by using Pd 0 , followed by cyclization with PyBOP.
  • the resulting crude peptides were purified by RP-HPLC and characterized by ESI-MS.
  • Cyclic AMP accumulation was determined in subconfluent cultures of HEK293 cells stably expressing GLP-IR in the presence of the phosphodiesterase inhibitor 3- isobutyl-1-methylxanthine (IBMX)
  • IBMX 3- isobutyl-1-methylxanthine
  • DPP-IV [4] A peptide (100 ⁇ M) was incubated with DPP-IV (0.2 ng/niL) at 37 0 C in Tris buffer (25 mM, pH 8.0). At the time of 0, 2, 4, 6, 9, and 24 h, an aliquot (100 ⁇ L) was taken out and quenched with aqueous TFA (10%, 20 ⁇ L) and analyzed by RP- HPLC (10-90% CH 3 CN in water (0.1% TFA) over 40 min; flow rate, 1.0 mL/min; C18-bonded Zorbax column, 4.6x250 mm).
  • NEP 24.11 [5]: A peptide (50 ⁇ M) was incubated with NEP 24.11 (20 ng/niL) at 37 0 C in HEPES buffer (50 niM, pH 7.4, 50 niM NaCl). At the time of 0 and 24 h, an aliquot (100 ⁇ L) was taken out and quenched with aqueous TFA (10%, 20 ⁇ L) and analyzed by RP-HPLC.
  • Figure 3 is a chart that shows receptor activation by cyclic GLP-I analogues.
  • Figure 4 are HPLC chromatograms of a bicyclic GLP-I analogue after incubation with DPP-IV and NEP,
  • Figure 5 is the synthesis of a PET imaging agent by using a bicyclic GLP-I analogue.
  • Figure 6 is a graph that shows the binding of L-GLP-I, D-GLP-I, EM2196 and EM2198 to various organs.
  • Figure 7 is a PET image of the binding of EM2198 in a mouse.
  • Figure 8 is a PET image of the binding of EM2198 with the addition of Extendin-4 in a mouse.
  • Figure 9 are graphs that show a comparison of contrast in pancreatic, liver and kidney microPET scans.
  • Figure 10 is a transaxial PET/CT images at 30 min p.i. of EM2198 with or without exendin-4 blocking. Based on the quantitative analysis of microPET/CT images, EM2198 showed the highest pancreas contrast. Its GLP-IR binding specificity was confirmed by a blocking imaging study using exendin-4.
  • Figure 11 is a graph that shows that EM2198 specifically target the GLP-IR in pancreas. Quantitative analysis based on microPET/CT 0-60 min dynamic scans, animal model: Normal BABL/c mice at 6-8 wks. Based on the quantitative analysis of microPET/CT images, EM2198 showed the highest pancreas contrast. Its GLP-IR binding specificity was confirmed by a blocking imaging study using exendin-4.
  • Figure 12 is a graph that illustrates binding of the bicyclic GLP-I analogue (EM2198) and Exendin-4.
  • the present invention also provides a novel Exendin-4 biomarker.
  • novel cyclic Exendin-4 analogues (as well as noncyclic Exendin-4 analogues) were designed and synthesized and proved to show significantly improved stability against enzymatic degradation.
  • the cyclic structure enhanced the potency of the cyclic Exendin-4 analogues that allowed clear detection of pancreatic beta-cells.
  • the Exendin-4 sequence includes: His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu- Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Pro- Ser-NH 2 SEQ ID No: 2 as well as variations and modifications thereof.
  • Exendin-4 sequence may be substituted and modified to be similar (having 75, 80, 85, 90, 95, 98 or 99% homology) to the Exendin-4 sequence above.
  • the novel cyclic Exendin-4 analogues can be labeled with proteins, radionuclides, fluorescent labels, metals, chromogenic agents, enzymes and other agents that enhace its use as an imaging agents.
  • radionuclides include, e.g., 18 F, 68 Ga, 60/61/62/64 c u , 89 Zr, 86 Y, 124 I, 99m Tc, 111 In, 67 Ga, 125 I, 123 I, 177 Lu, 166 Ho, and 153 Sm.
  • the imaging agent further includes at least one of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
  • the agent further comprises at least one of an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an antimitotic agent, an anthracycline, toxin, and an apoptotic agent.
  • the imaging agent may include or more labels that make the agent detectable by positron emission tomography (PET), single photon emission computed tomography (SPECT), radioscintigraphy, magnetic resonance imaging (MRI) and computed tomography (CT scan).
  • PET positron emission tomography
  • SPECT single photon emission computed tomography
  • MRI magnetic resonance imaging
  • CT scan computed tomography
  • the agents disclosed herein have been found to have increased resistance to proteolytic cleavage by dipeptidyl peptidase-IV (DPP-IV), neutral endopeptidase (NEP), or both.
  • Exendin-4 found in the venom of the GiIa Monster (Heloderma suspetum), is also an agonist for the GLP-I receptor (GLP-IR). It is resistant to DPP IV digestion and, unlike GLP-I, it can be truncated by 8 amino acid residues at its N-terminus without losing receptor affinity. However, the loss of the first 2-8 amino acid residues results in the generation of antagonists.
  • the N-terminal region of GLP-I and exendin-4 are almost identical, a significant difference being the second amino acid residue, alanine in GLP-I and glycine in Exendin-4, which gives Exendin-4 its resistance to DPP-IV digestion.
  • Exendin-4 has an extra 9 amino acid residues at its C-terminus which have been shown to form a 'Trp-cage' by NMR. NMR analysis of Exendin-4 also shows that the central region (amino acid residues 10-30) is helical in structure. Interestingly GLP-I and Exendin-4 only share 8 amino acid residues in this region but since they lie on the same face of the ⁇ -helix, we postulate that this face of the helix interacts with the receptor.
  • GLP-I receptor has been cloned and is a member of the 'family B' G protein- coupled receptors (GPCRs). Other members of this family include receptors for glucagon, calcitonin, glucose-dependent insulinotropic polypeptide and vasoactive intestinal peptide. It is known that the large amino terminal domain that characterizes the 'family B' GPCRs plays a key role in ligand binding. However, the amino terminus is not entirely sufficient to bind the ligand and regions in the exctracellular loops and/or transmembrane helices are also believed to provide additional interactions.
  • Exendin-4 is another ideal candidate to be employed for non-invasive imaging of the beta-cells.
  • molecular imaging is gaining significant attention in the basic biomedical sciences and in clinical research and practice.
  • non-invasive imaging techniques are revolutionizing the understanding of diseases at the cellular and molecular levels.
  • the ability to non-invasively visualize pancreatic beta-cells would greatly facilitate the development of new methods in the prevention and treatment of diabetes.
  • Conventional magnetic resonance imaging (MRI) and computed tomography (CT) can be used to delineate the location of the pancreas in a subject at a spatial resolution of ⁇ 100 ⁇ m.
  • pancreas is a highly vascularized soft organ and the islets only represent 2-3 % of the pancreatic tissues.
  • imaging or contrast agents that recognize the scarcely dispensed beta-cells within pancreas and are responsive to their biological functions, must be developed.
  • tomographic nuclear imaging approaches especially positron emission tomography (PET) have demonstrated their significant importance and promising potential in applications of molecular imaging probes due to the superior sensitivity and specificity in diverse subjects, and the ability to quantitatively analyze the regions of interest.
  • Exendin-4 a 39 amino acid peptide originally isolated from the oral secretions of the lizard Heloderma suspectum, has been shown to share certain activities with glucagon-like-peptide-1 (GLP-I), a 30 amino acid peptide.
  • GLP-I glucagon-like-peptide-1
  • Exendin-4 play a critical role in the function of beta-cells.
  • Exendin-4 includes the sequence: DLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS- NH 2 SEQ ID No: 3 and also may include Exendin-4 constructs (1-39, 4-39, 9-39, 1-30, 4-30, 9- 30 or any variation thereof). Other Exendin-4 constructs may be used as well as other variations and mutations of Exendin.
  • Exendin-4 His Ser Asp GIy Thr Phe Thr Ser Asp Leu Ser Lys GIn Met GIu GIu GIu Ala VaI Arg Leu Phe lie GIu Trp Leu Lys Asn GIy GIy Pro Ser Ser GIy Ala Pro Pro Pro Ser NH 2 SEQ ID No: 3.
  • the present invention includes Exendin constructs with similar modifications, substations, and construct sequences as illustrated herein for glucagon-like peptide- 1 (GLP-I).
  • the present invention discloses an imaging composition of an cyclic analog uses a linker to connect a portion of a peptide or protein that binds specifically to the GLP-I receptor (GLP-IR) and the cyclic analog has one or more conformational restrictions including, but not limited to, lactam bridges, disulfide bridges, hydrocarbon bridges, and their combinations, salts and derivatives thereof wherein the cyclic analog is more stable than a non-cyclic analog when incubated in the presence of enzymes that degrade GLP-I and have an increased serum half- live to an imaging molecule.
  • GLP-IR GLP-I receptor
  • composition of the instant invention can be divided into three regions, a GLP-lR-binding peptide, a linker (or spacer), and a metal chelator (cyclic or linear).
  • linker molecule may be 6-aminohexanoyl-cysteine or the thiol of the cysteine to make a covalent bond with DOTA (or NOTA)-maleimide.
  • DOTA or NOTA
  • other linkers may be used.
  • the linker may include alpha-amino acids, omega-amino acids (e.g., beta- alanine, 4-aminobutyric acid, 5 -amino valeric acid, 6-aminohexanoic acid), A- aminocyclohexanecarboxylic acid, 3-aminocyclohexanecarboxylic acid, 2- aminocyclohexanecarboxylic acid, 3-aminocyclopentanecarboxylic acid, 2- aminocyclopentanecarboxylic acid, 4-aminobenzoic acid, 3-aminobenzoic acid, 2-aminobenzoic acid, poly(ethylene glycol).
  • omega-amino acids e.g., beta- alanine, 4-aminobutyric acid, 5 -amino valeric acid, 6-aminohexanoic acid
  • A- aminocyclohexanecarboxylic acid 3-aminocyclohexanecarboxylic acid
  • the metal chelator may be HYNIC, Bolton-Hunter moiety, cross- bridged DOTA and TETA (CB-DO2A and CB-TE2A) NOTA, and DTPA.
  • the present invention also provide different combination of peptide, linkers and metal chelators
  • compositions of the invention can be used to achieve methods of the invention.
  • the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), "including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.
  • A, B, C, or combinations thereof refers to all permutations and combinations of the listed items preceding the term.
  • A, B, C, or combinations thereof is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB.
  • expressly included are combinations that contain repeats of one or more item or term, such as BB, AAA, MB, BBC, AAABCCCC, CBBAAA, CABABB, and so forth.
  • the skilled artisan will understand that typically there is no limit on the number of items or terms in any combination, unless otherwise apparent from the context.
  • words of approximation such as, without limitation, "about”, “substantial” or “substantially” refers to a condition that when so modified is understood to not necessarily be absolute or perfect but would be considered close enough to those of ordinary skill in the art to warrant designating the condition as being present.
  • the extent to which the description may vary will depend on how great a change can be instituted and still have one of ordinary skilled in the art recognize the modified feature as still having the required characteristics and capabilities of the unmodified feature.
  • a numerical value herein that is modified by a word of approximation such as "about” may vary from the stated value by at least ⁇ 1, 2, 3, 4, 5, 6, 7, 10, 12 or 15%.
  • compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

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Abstract

Cette invention concerne des compositions, des méthodes d'utilisation et des méthodes de fabrication d'un analogue peptidique cyclique utilisé comme agent en imagerie et comprenant au moins des portions d'un peptide ou d'une protéine qui se fixe spécifiquement au récepteur GLP-1 (GLP-1R); l'analogue cyclique comporte une ou plusieurs restrictions de conformation, notamment des liaisons lactame, des liaisons bisulfure, des liaisons hydrocarbonées, et leurs combinaisons, sels et dérivés; l'analogue cyclique est plus stable qu'un analogue non cyclique une fois incubé en présence d'enzymes dégradant le récepteur GLP-1, et présente une demi-vie sérique accrue, l'analogue cyclique comprenant au moins une portion d'un peptide de GLP-1 ou au moins une portion d'un peptide d'extension, ses sels, dérivés ou combinaisons.
PCT/US2010/030103 2009-04-06 2010-04-06 Analogues peptidiques cycliques utilisés en imagerie non invasive des cellules bêta du pancréas Ceased WO2010118034A2 (fr)

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UA116217C2 (uk) 2012-10-09 2018-02-26 Санофі Пептидна сполука як подвійний агоніст рецепторів glp1-1 та глюкагону
EP2927239B1 (fr) * 2012-11-30 2019-09-04 Kyoto University Polypeptide et procédé d'imagerie
MA38276B1 (fr) 2012-12-21 2018-03-30 Sanofi Sa Dérivés de l'exendine 4 pour l’utilisation dans le traitement des troubles du syndrome metabolique, y compris le diabete et l'obesite, ainsi que la reduction de l'apport alimentaire excessif.
TW201609795A (zh) 2013-12-13 2016-03-16 賽諾菲公司 作為雙重ｇｌｐ－１／ｇｉｐ受體促效劑的艾塞那肽－４（ｅｘｅｎｄｉｎ－４）胜肽類似物
WO2015086729A1 (fr) 2013-12-13 2015-06-18 Sanofi Agonistes mixtes des récepteurs glp-1/gip
EP3080149A1 (fr) 2013-12-13 2016-10-19 Sanofi Agonistes mixtes des récepteurs du glp-1/glucagon
TW201609796A (zh) 2013-12-13 2016-03-16 賽諾菲公司 非醯化之艾塞那肽－４（ｅｘｅｎｄｉｎ－４）胜肽類似物
TW201625669A (zh) 2014-04-07 2016-07-16 賽諾菲公司 衍生自艾塞那肽－４（Ｅｘｅｎｄｉｎ－４）之肽類雙重ＧＬＰ－１／升糖素受體促效劑
TW201625668A (zh) 2014-04-07 2016-07-16 賽諾菲公司 作為胜肽性雙重ｇｌｐ－１／昇糖素受體激動劑之艾塞那肽－４衍生物
TW201625670A (zh) 2014-04-07 2016-07-16 賽諾菲公司 衍生自ｅｘｅｎｄｉｎ－４之雙重ｇｌｐ－１／升糖素受體促效劑
US9932381B2 (en) 2014-06-18 2018-04-03 Sanofi Exendin-4 derivatives as selective glucagon receptor agonists
AR105319A1 (es) 2015-06-05 2017-09-27 Sanofi Sa Profármacos que comprenden un conjugado agonista dual de glp-1 / glucagón conector ácido hialurónico
AR105284A1 (es) 2015-07-10 2017-09-20 Sanofi Sa Derivados de exendina-4 como agonistas peptídicos duales específicos de los receptores de glp-1 / glucagón
CN109078197A (zh) * 2018-09-12 2018-12-25 江苏省原子医学研究所 一种新型胃排空测定方法

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DE102004043153B4 (de) * 2004-09-03 2013-11-21 Philipps-Universität Marburg Erfindung betreffend GLP-1 und Exendin
ES2495741T3 (es) * 2006-04-20 2014-09-17 Amgen, Inc Compuestos de GLP-1
US7816324B2 (en) * 2007-03-13 2010-10-19 Board Of Regents, The University Of Texas System Composition and method for the treatment of diseases affected by a peptide receptor
JP6108659B2 (ja) * 2008-06-17 2017-04-05 インディアナ ユニバーシティー リサーチ アンド テクノロジー コーポレーションＩｎｄｉａｎａ Ｕｎｉｖｅｒｓｉｔｙ Ｒｅｓｅａｒｃｈ Ａｎｄ Ｔｅｃｈｎｏｌｏｇｙ Ｃｏｒｐｏｒａｔｉｏｎ 代謝疾患および肥満の治療のためのｇｉｐに基づいた混合アゴニスト

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Publication number Priority date Publication date Assignee Title
EP2578599A1 (fr) 2011-10-07 2013-04-10 LanthioPep B.V. Analogues cycliques de GLP-1 et peptides de type GLP-1
WO2013051938A2 (fr) 2011-10-07 2013-04-11 Lanthiopep B.V. Analogues peptidiques cycliques de glp-1 ou de peptides apparentés à glp-1 et leurs utilisations

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